Fig. 2 | Nature Communications

Fig. 2

From: Reversible histone glycation is associated with disease-related changes in chromatin architecture

Fig. 2

Histone H3 and H4 are prime targets for glycation in vitro and in cellulo. a Glycation of free histones. Recombinant purified histones (indicated in the coomassie brilliant blue stained SDS-PAGE) were incubated with increasing concentrations of MGO (0.1, 0.5 or 1 mM, corresponding to 4:1, 1:1 or 1:2 sites:MGO stoichiometry, respectively) for 12 h at 37 °C. Analysis was performed by SDS-PAGE followed by western blot with anti-MGO. b Reconstituted nucleosomes were treated with 0, 0.5, 5 or 100 mM of MGO (corresponding to 1:2, 1:20 or 1:400 sites:MGO stoichiometry, respectively) for 72 h at 37 °C, after which the nucleosomes were analyzed as described for free histones. c Histone glycation in 293T cells after 0.5 mM MGO treatment for 24 h. Histones were extracted by high salt and analyzed by coomassie or western blotted with the indicated antibodies. d 293T cells were incubated with 0, 0.25, 0.5 or 1 mM of MGO for 12 h at 37 °C, after which histones were extracted with high salt and analyzed by western blot with the indicated histone PTM antibodies